单糖紫外衍生方法的改进与应用  

Improvement and Application of An Ultraviolet Derivative Method for Monosaccharides

作　　者：郭怀忠[1,2,3] 李胜男[1,2] 吴芳[1,2] 杨飞飞[1,2] 李玉青[1] 

机构地区：[1]河北大学药学院,河北保定071002 [2]河北省药物质量分析控制重点实验室,河北保定071002 [3]药物化学与分子诊断教育部重点实验室,河北保定071002

出　　处：《中国实验方剂学杂志》2017年第2期32-36,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(21575033);河北省自然科学基金项目(H2016201221)

摘　　要：目的:建立一种通过在1-苯基-3-甲基-5-吡唑啉酮(PMP)和对氨基苯甲酸(PABA)衍生单糖反应中加入硼砂,以提高单糖衍生效率的方法。方法:实验考察了硼砂的加入对PMP衍生8种常见醛糖和PABA衍生酮糖(以果糖为例)衍生效率的影响,优化了硼砂溶液浓度、温度及时间等衍生条件,采用毛细管区带电泳(CZE)进行方法学考察,并将改进的方法应用于中药南沙参的单糖组成测定。结果:在衍生过程中加入硼砂均可一定程度提高PMP和PABA对单糖的衍生效率,其中以葡萄糖最为显著,可提高约1.2倍。优化的PMP衍生醛糖的条件为:硼砂溶液浓度6.86 mmol·L^(-1),衍生温度70℃,衍生时间30 min。优化的PABA衍生果糖的条件为:硼砂溶液浓度20.43 mmol·L^(-1),衍生温度70℃,衍生时间60 min。在测定南沙参单糖组成时,改进的方法使一些含量较低的、原方法未发现的单糖被检测出来。结论:硼砂的加入可改变单糖构象平衡,链状构象占比增加,从而提高了单糖衍生效率。该方法准确、灵敏、操作简便,可为改善中药糖类成分分析及构效关系研究提供支持。Objective： A monosaccharide derivative method with improved efficiency was developed through the addition of borax in the process of 1-penyl-3-methyl-5-pyrazolone（ PMP） and p-aminobenzoic acid（ PABA） derivatization. Method： The effect of borax on the derivative efficiency of 8 common aldoses with PMP,and ketose（ fructose） with PABA was investigated. Derivative conditions such as the concentration of borax solution,temperature and time were optimized by capillary zone electrophoresis（ CZE） method. The improved method was applied to the determination of monosaccharide composition of Adenophorae Radix polysaccharide.Result： The results showed that the addition of borax could enhance both the PMP and PABA derivative efficiencies during the monosaccharide derivatization,and the most significant improved efficiency belonged to glucose,which could be increased roughly 1. 2-fold. The optimized conditions of aldoses derivatization with PMP were as follows：the concentration of borax solution was 6. 86 mmol·L^-1,the temperature was 70 ℃,and the time was 30 min; the optimized conditions of ketose derivatization with PABA were as follows： the concentration of borax solution was20. 43 mmol·L^-1,the temperature was 70 ℃,and the time was 60 min. The application of the improved method to the determination of monosaccharide composition of Adenophorae Radix polysaccharide showed monosaccharide with lower content could be detected by the improved method while the original one failed. Conclusion： Therefore,it is an useful approach to improve the monosaccharide derivative efficiency and detection sensitivity by adding the borax to change the balance of conformations of monosaccharide,which makes the proportion of chain structure increased.The method is accurate,sensitive and simple,and provides technical support for the monosaccharide composition analysis and the evaluation of structure-activity relationship of polysaccharide from traditional Chinese medicines.

关 键 词：衍生效率 硼砂 醛糖 果糖 毛细管区带电泳 

分 类 号：R284.1[医药卫生—中药学]